Glycosaminoglycans from two human malignant mesothelioma cell lines: determination, distribution, and effect of platelet-derived growth factor on their synthesis

Abstract

The synthesis and distribution of glycosaminoglycans (GAGs) were studied in two human malignant mesothelioma cell lines: one with fibroblast-like morphology and the other with epithelial differentiation. Analyses using highly sensitive high-pressure liquid chromatography techniques and agarose gel electrophoresis showed that these cells produce not only hyaluronan (HA) but also galactosaminoglycans (GalAGs, chondroitin sulfate and (or) dermatan sulfate) and heparan sulfate (HS). In both cell lines most of the HA (87-90%) and GalAGs (57-66%) are secreted into the extracellular matrix. Although HS is mainly bound to the cell surface in fibroblast-differentiated cells (75%), in epithelial type cells only 40% occurs in the cell-associated fraction. The amounts of secreted GAGs are 6- to 8-fold higher in epithelial than in fibroblast-like mesothelioma cultures. In cells with the fibroblast phenotype, the beta-homodimer of platelet-derived growth factor (PDGF) in a concentration of 1.5 ng/mL stimulates HA and GalAG synthesis 5-fold and that of HS 10-fold, whereas higher concentrations suppress this stimulatory effect. The stimulatory effect, observed at low concentrations of this growth factor, was completely blocked by the addition of antibodies against this factor. In epithelially differentiated cells, the production of all GAGs was suppressed after addition of this factor, even at low concentrations. We therefore suggest that mesothelioma cells can produce GAGs, the synthesis of which is dependent on the presence and concentration of PDGF beta-homodimer. The differences between the two cell lines regarding the effect of this growth factor on GAG synthesis indicates that the regulation of this synthesis is complex, other factors also being important.

Regulation of matrix metalloprotease activity in malignant mesothelioma cell lines by growth factors

“…14 17 Furthermore, it has been shown that a number of growth factors stimulate the synthesis of the extracellular matrix molecule hyaluronan and proteoglycans in malignant mesothelioma cells. 25 It has been indicated that increased production of hyaluronan by tumour cells or tumour associated fibroblasts is correlated with cell migration and metastasis. 26 In our study we have shown that these growth factors also increase the secretion of MMP-9 and MMP-3 in malignant mesothelioma cells.…”

Synthesis and distribution of glycosaminoglycans in human leukemic B‐ and T‐cells and monocytes studied using specific enzymic treatments and high‐performance liquid chromatography

“…Proteoglycan synthesis by normal lymphocytes and monocytes has also been found to be affected by mitogenic activation of cells, indicating that the synthesis and secretion of PGs by activated leukocytes may well be related to cellular functions (Wilson and Rider, 1991;Barthold et al, 1989). It has recently been reported (Tzanakakis et al, 1995Syrokou et al, 1999) that mitogenic substances affect both synthesis and distribution of GAGs/PGs on human malignant mesothelioma cells, independently of their effect on cell proliferation. This study was therefore undertaken to demonstrate the type of glycosaminoglycans produced from leukemic T-and B-cells and monocytic origin, and their distribution among cell membrane and extracellular matrix as compared with those produced by normal PBML.…”

Hyaluronan Determination: Biological Significance & Analytical Tools

“…This method has been applied to the routine determination of HA in pleural fluid from patients with malignant mesothelioma [108,109], of HA in aqueous humor of patients with exfoliation syndrome [110], in human normal and aneurysmal abdominal aortas [95], and in pancreatic carcinoma [111]. Furthermore, HA secreted in cell cultures was easily determined after treatment of cells with [6-3 H]-D-glucosamine and HPLC analysis using a radiochemical detector [112][113][114].…”